Dietary and genetic manipulations of folate metabolism differentially affect neocortical functions in mice.

Converging evidence suggests that folate-mediated one-carbon metabolism may modulate cognitive functioning throughout the lifespan, but few studies have directly tested this hypothesis. This study examined the separate and combined effects of dietary and genetic manipulations of folate metabolism on neocortical functions in mice, modeling a common genetic variant in the MTHFD1 gene in humans. Mutant (Mthfd1(gt/+)) and wildtype (WT) male mice were assigned to a folate sufficient or deficient diet at weaning and continued on these diets throughout testing on a series of visual attention tasks adapted from the 5-choice serial reaction time task. WT mice on a deficient diet exhibited impulsive responding immediately following a change in task parameters that increased demands on attention and impulse control, and on trials following an error. This pattern of findings indicates a heightened affective response to stress and/or an inability to regulate negative emotions. In contrast, Mthfd1(gt/+) mice (regardless of diet) exhibited attentional dysfunction and a blunted affective response to committing an error. The Mthfd1(gt/+) mice also showed significantly decreased expression levels for genes encoding choline dehydrogenase and the alpha 7 nicotinic cholinergic receptor. The effects of the MTHFD1 mutation were less pronounced when combined with a deficient diet, suggesting a compensatory mechanism to the combined genetic and dietary perturbation of folate metabolism. These data demonstrate that common alterations in folate metabolism can produce functionally distinct cognitive and affective changes, and highlight the importance of considering genotype when making dietary folate recommendations.

A mouse model of fragile X syndrome exhibits heightened arousal and/or emotion following errors or reversal of contingencies.

This study was designed to further assess cognitive and affective functioning in a mouse model of Fragile X syndrome (FXS), the Fmr1(tm1Cgr) or Fmr1 "knockout" (KO) mouse. Male KO mice and wild-type littermate controls were tested on learning set and reversal learning tasks. The KO mice were not impaired in associative learning, transfer of learning, or reversal learning, based on measures of learning rate. Analyses of videotapes of the reversal learning task revealed that both groups of mice exhibited higher levels of activity and wall-climbing during the initial sessions of the task than during the final sessions, a pattern also seen for trials following an error relative to those following a correct response. Notably, the increase in both behavioral measures seen early in the task was significantly more pronounced for the KO mice than for controls, as was the error-induced increase in activity level. This pattern of effects suggests that the KO mice reacted more strongly than controls to the reversal of contingencies and pronounced drop in reinforcement rate, and to errors in general. This pattern of effects is consistent with the heightened emotional reactivity frequently described for humans with FXS.

Monitoring weight daily blocks the freshman weight gain: a model for combating the epidemic of obesity.

BACKGROUND: The Tissue Monitoring System (TMS) is an algorithm that estimates changes in body tissue from a series of daily weight measures. It is intended to provide people with a feedback of changes in their tissue weight so they may have a basis for estimating how much they would have to change their intake or expenditure to maintain their weight at a prescribed level. We tested the effectiveness of the TMS to prevent freshmen from gaining weight during their first semester in college. METHODS: In two similar but independent studies (Fall 2002, 2003), female freshmen college students were given analog bathroom scales and instructed to weigh themselves each morning immediately after rising from bed, then e-mail their weight to our staff. After 7 days, a linear function was performed on the most recent 7 days of the weight-day function for each participant. In the first study, the slope of this function was e-mailed back to the participants. In the second study, the difference between last point and the original weight was determined, using linear regression techniques, converted to calories, and the information was e-mailed back to the participants. Control participants in both studies were weighed at the beginning and the end of the semester. RESULTS: The untreated controls gained 3.1+/-0.51 kg and 2.0+/-0.65 kg, respectively (P<0.01 for both studies), whereas weight gain of the experimental groups was 0.1+/-0.99 kg and -0.82+/-0.56 kg, values that were not significantly different than zero. CONCLUSIONS: The TMS appears to be an effective technique to help female college freshmen resist gaining weight in an environment that is conducive to weight gain. These results suggest that the TMS may be a useful method to help curb the slow increase in the prevalence of overweight and obesity that is characteristic of all industrialized societies.

Attentional dysfunction, impulsivity, and resistance to change in a mouse model of fragile X syndrome.

On a series of attention tasks, male mice with a mutation targeted to the fragile X mental retardation 1 (Fmrl) gene (Fmrl knockout [KO] mice) committed a higher rate of premature responses than wild-type littermates, with the largest differences seen when task contingencies changed. This finding indicates impaired inhibitory control, particularly during times of stress or arousal. The KO mice also committed a higher rate of inaccurate responses than controls, particularly during the final third of each daily test session, indicating impaired sustained attention. In the selective attention task, the unpredictable presentation of potent olfactory distractors produced a generalized disruption in the performance of the KO mice, whereas for controls, the disruption produced by the distractors was temporally limited. Finally, the attentional disruption seen following an error was more pronounced for the KO mice than for controls, further implicating impaired regulation of arousal and/or negative affect. The present study provides the first evidence that the Fmrl KO mouse is impaired in inhibitory control, attention, and arousal regulation, hallmark areas of dysfunction in fragile X syndrome. The resistance to change also seen in these mice provides a behavioral index for studying the autistic features of this disorder.

The freshman weight gain: a model for the study of the epidemic of obesity.

OBJECTIVE: The objective of this study was to quantify the weight gain of freshmen during their first 12 weeks at Cornell University. In addition, students completed questionnaires that revealed particular behaviors and activities that were associated with weight gain. DESIGN: Serial, correlational study. SUBJECTS: A total of 68 freshmen from Cornell University. MEASUREMENT: A total of 60 students were weighed during the first week of the semester, then again 12 weeks later. They were also given a questionnaire to complete concerning their behavior during the previous 12 weeks. RESULTS: After adjusting for clothing weights, the mean weight gain of the freshmen was 1.9+/-2.4 kg, a value significantly different from 0. Two regression models generated from the questionnaire were fitted to the weight gain. The first linear regression model (Model 1) accounted for 58% of the variance and indicated that eating in the 'all-you-can-eat' dining halls accounted for 20% of the variance in weight gain. Snacking and eating high-fat 'junk food' accounted for anther 20%. When initial weight was used as a covariate (Model 2), the consumption of junk foods, meal frequency and number of snacks accounted for 47% of the variance. CONCLUSION: The study clearly demonstrated that significant weight gain during first semester college is a real phenomenon and can be attributed to tangible environmental stimuli. The weight gain is considerably greater than that observed in the population and may be useful as a model to test various techniques that may reduce or reverse the 'epidemic' of obesity observed in the general population.

The effect of microgravity and space flight on the chemical senses.

The effect of space flight and microgravity on the chemical senses is reviewed. Skylab-4 and Soyuz 30-31 studies revealed changes in taste thresholds while no effect was found in a Canadian investigation (41-G) and conflicting results were obtained on another Soyuz mission. Two simulated microgravity studies found no effect on taste or smell sensitivity; while 5 other studies found an effect. Microgravity induces physiological changes including an upward shift of body fluids toward the head, which may lead to an attenuation of the olfactory component in the flavor of foods. Chemosensory changes may also relate to space sickness, Shuttle atmosphere, stress, radiation, and psychological factors.

Recovery of associative function following early amygdala lesions in rats.

Adult rats with amygdala lesions made at either Postnatal Day (PND) 10 or PND40 were tested on a series of reversal tasks that tap the ability to form stimulus-reward associations. PND40 rats were significantly impaired relative to both controls and PND10 rats on learning rate of the original discrimination and subsequent reversals. Analyses of discrete learning phases revealed that the impairment was specific to the postchance phase. The PND10 group was not impaired relative to controls on any measure. These results confirm prior findings that amygdala lesions sustained in adulthood impair the formation of stimulus-reward associations. They also demonstrate that substantial sparing or recovery of function is possible when the lesion is made during early development. Furthermore, the findings support the view that behavioral recovery may be more likely if the lesion is sustained near the time of peak synaptogenesis.

Early lead exposure produces lasting changes in sustained attention, response initiation, and reactivity to errors.

The present study tested the hypothesis that early lead (Pb) exposure causes lasting attentional dysfunction. Long-Evans dams were fed Pb-adulterated water during gestation and/or lactation; the offspring were tested as adults. The results of a visual discrimination task revealed no Pb effects on learning rate or information-processing speed. However, lasting effects of the early Pb exposure were seen in the subsequent vigilance tasks, particularly in the final task in which onset of the visual cue and cue duration varied randomly across trials. Exposure during both gestation and lactation impaired response initiation. In addition, animals exposed to Pb during lactation only or lactation+gestation committed significantly more omission errors than controls under two specific conditions: (1) trials in which a delay was imposed prior to cue presentation and (2) trials that followed an incorrect response. The pattern of treatment differences indicated that early Pb exposure produced lasting impairment of sustained attention and increased reactivity to errors. Both effects may contribute to the cognitive impairment, problematic classroom behaviors, and increased delinquency associated with early Pb exposure in children. These findings also demonstrate that the developmental timing of the exposure determines the pattern of effects. Thus, conclusions regarding whether or not a particular cognitive or affective function is impaired or spared by early Pb exposure must be limited to the specific timing and intensity of exposure.

Prenatal cocaine exposure impairs selective attention: evidence from serial reversal and extradimensional shift tasks.

This study assessed the effects of prenatal cocaine exposure on cognitive functioning, using an intravenous (IV) rodent model that closely mimics the pharmacokinetics seen in humans after smoking or IV injection and that avoids maternal stress and undernutrition. Cocaine-exposed males were significantly impaired on a 3-choice, but not 2-choice, olfactory serial reversal learning task. Both male and female cocaine-exposed rats were significantly impaired on extradimensional shift tasks that required shifting from olfactory to spatial cues; however, they showed no impairment when required to shift from spatial to olfactory cues. In-depth analyses of discrete learning phases implicated deficient selective attention as the basis of impairment in both tasks. These data provide clear evidence that prenatal cocaine exposure produces long-lasting cognitive dysfunction, but they also underscore the specificity of the impairment.

Effects of chronic lead exposure on learning and reaction time in a visual discrimination task.

Long-Evans rats exposed chronically to lead (Pb) acetate (0, 75, or 300 ppm) were tested as adults on an automated, three-choice visual discrimination task as part of a larger study designed to elucidate the cognitive effects of developmental Pb exposure. Median adult BPb levels for the groups were <5, 20, and 36 microgram/dl. The pattern of results suggested a linear effect, with increasing lead dose producing progressively slower learning and an increased incidence of "impaired" individuals. This latter measure proved to be slightly more sensitive than the former, suggesting individual differences in susceptibility to Pb neurotoxicity. Additional analyses revealed that the impairing effect of Pb was seen in both the chance and post-chance learning phases, indicating that the deficit was not limited to (but could include) attentional function. Reaction time on incorrect trials was reduced in the 300-ppm group, whereas no Pb effect was seen for correct trials. The present findings suggest that chronic developmental Pb exposure produces an associative deficit as well as a tendency to respond rapidly, but does not affect information-processing speed.

Enduring effects of early lead exposure: evidence for a specific deficit in associative ability.

Long-Evans dams were exposed to Pb acetate in the drinking water during both gestation and lactation, or lactation only. This report presents the results of an automated, olfactory, serial reversal task administered to the adult offspring. Although overall learning rate was not significantly affected by Pb exposure, analyses of specific phases of the learning process revealed that all three exposed groups required significantly more trials than controls to reach criterion from the point at which perseverative responding to the previously correct cue ended. These in-depth analyses revealed that the reversal learning impairment of the Pb-exposed animals was not due to a deficit in inhibiting responses to the previously correct cue, the mechanism commonly assumed to underlie impaired reversal learning. Instead, the analyses revealed that two other independent Pb effects were responsible for the prolonged postperseverative learning period: a response bias and an impaired ability to associate cues and/or actions with affective consequences. The contribution of these two factors varied as a function of the timing and intensity of the Pb exposure. It is hypothesized that the Pb-induced associative deficit may reflect lasting damage to the amygdala and/or nucleus accumbens, which comprise a system thought to modulate the process by which environmental cues acquire affective significance.

The promotion effect of anorectic drugs on aflatoxin B(1)-induced hepatic preneoplastic foci.

The ability of three extensively used anorectic drugs, namely fenfluramine (FN), fluoxetine (FX) and amphetamine (AM), to alter the development of aflatoxin B(1) (AFB(1))-induced gamma-glutamyl-positive (GGT(+)) preneoplastic liver foci was investigated in 135 male weanling F344 rats. Following AFB(1) administration, 15 rats were killed, while the rest were divided into four groups and fed diets containing either FN, FX, AM or control diet, with half of the animals in each group subsequently being killed at 4 weeks and half at 10 weeks. All three anorectic drugs as expected suppressed initial food intake, growth rate, body weight gain and food efficiency. They also tended to suppress body fat mass and to decrease plasma levels of T(3) and T(4). FN significantly (P < 0.05) increased GGT(+) foci number/cm(2) and number/cm(3), while FX significantly increased GGT(+) foci number/cm(2) and the volume fraction of foci. Histopathological staining also revealed that FN- and FX-treated animals had more serious morphological alterations in their liver tissue. In contrast, foci development was, if anything, suppressed by AM feeding. These results indicate that serotoninergic drugs (FN and FX), as opposed to dopaminergic drugs (AM), may have tumor promoter activity, at least for liver tissue.

Effects of dietary mixtures of amino acids on fetal growth and maternal and fetal amino acid pools in experimental maternal phenylketonuria.

BACKGROUND: Branched-chain amino acids have been reported to improve fetal brain development in a rat model in which maternal phenylketonuria (PKU) is induced by the inclusion of an inhibitor of phenylalanine hydroxylase, DL-p-chlorophenylalanine, and L-phenylalanine in the diet. OBJECTIVE: We studied whether a dietary mixture of several large neutral amino acids (LNAAs) would improve fetal brain growth and normalize the fetal brain amino acid profile in a rat model of maternal PKU induced by DL-alpha-methylphenylalanine (AMPhe). DESIGN: Long-Evans rats were fed a basal diet or a similar diet containing 0.5% AMPhe + 3.0% L-phenylalanine (AMPhe + Phe diet) from day 11 until day 20 of gestation in experiments to test various mixtures of LNAAs. Maternal weight gains and food intakes to day 20, fetal body and brain weights at day 20, and fetal brain and fetal and maternal plasma amino acid concentrations at day 20 were measured. RESULTS: Concentrations of phenylalanine and tyrosine in fetal brain and in maternal and fetal plasma were higher and fetal brain weights were lower in rats fed the AMPhe + Phe diet than in rats fed the basal diet. However, fetal brain growth was higher and concentrations of phenylalanine and tyrosine in fetal brain and in maternal and fetal plasma were lower in rats fed the AMPhe + Phe diet plus LNAAs than in rats fed the diet containing AMPhe + Phe alone. CONCLUSION: LNAA supplementation of the diet improved fetal amino acid profiles and alleviated most, but not all, of the depression in fetal brain growth observed in this model of maternal PKU.

Enhanced thermogenesis during recovery from diet-induced weight gain in the rat.

The present study examined the contribution of energy expenditure to the recovery of body weight after a period of overfeeding. Three groups of 2-mo-old female rats (n = 24) were fed, respectively, a 10% (wt/wt)-fat diet (control), a 35% (wt/wt)-fat diet (high fat) or were force fed 130% of the control diet (tube fed). After 30 days, all groups received the control diet for 18 days of recovery. Both overfeeding protocols significantly increased weight above control levels. This difference disappeared after 7 days of recovery. Increases in resting oxygen consumption, serum 3,5,3'-triiodothyronine (T3) levels, and the thermogenic response to norepinephrine were observed at the end of overfeeding. Serum T3 and resting oxygen consumption returned to control levels by day 3 of recovery from overfeeding, whereas the thermogenic response to norepinephrine required 9 days to recover. Whereas total energy expenditure was not significantly elevated during overfeeding, significant increases of 9.6 and 13.9% were observed in the formerly high-fat and tube-fed animals, respectively, during recovery. These data indicate that changes in energy expenditure play an important role in maintaining the stability of body weight.

The relationship between body weight and mortality: a quantitative analysis of combined information from existing studies.

OBJECTIVE: To estimate the relationship between body mass index (BMI, kg/m2) and all-cause mortality with information from the published scientific literature. DESIGN: Meta-analysis using a hierarchical, mixed model. The analysis included random effects for information sources and fixed effects for factors that may modify the BMI-mortality relationship such as smoking, control for disease, and country of origin, which allowed combining information from diverse studies. MAIN OUTCOME MEASURES: Predicted probability of death over a given duration of follow-up plotted by BMI for sex-age cohorts of white race. RESULTS: An extensive search identified nineteen prospective cohort studies that met inclusion criteria. A U-shaped relationship between BMI and mortality was demonstrated for 50-year-old men followed for 30 years. Mortality risk increased with low and high BMI (< 23 or > 28) in groups of non-smokers without evidence of disease upon study entry. Limited information from studies of women indicated that, with 10 year follow-up, there was little relationship between BMI and mortality for (1) non-smokers and for (2) mixtures of smokers and non-smokers. CONCLUSION: This quantitative analysis of existing studies revealed increased mortality at moderately low BMI for white men comparable to that observed at extreme overweight, which does not appear to be due to smoking or existing disease. Attention to the health risks of underweight is needed, and body weight recommendations for optimum longevity need to be considered in light of these risks.

Malnutrition and the brain: changing concepts, changing concerns.

Our conceptions of how malnutrition endured early in life affects brain development have evolved considerably since the mid-1960s. At that time, it was feared that malnutrition endured during certain sensitive periods in early development would produce irreversible brain damage possibly resulting in mental retardation and an impairment in brain function. We now know that most of the alterations in the growth of various brain structures eventually recover (to some extent), although permanent alterations in the hippocampus and cerebellum remain. However, recent neuropharmacological research has revealed long-lasting, if not permanent, changes in brain neural receptor function resulting from an early episode of malnutrition. These more recent findings indicate that the kinds of behaviors and cognitive functions impaired by malnutrition may be more related to emotional responses to stressful events than to cognitive deficits per se, the age range of vulnerability to these long-term effects of malnutrition may be much greater than we had suspected and the minimal amount of malnutrition (hunger) necessary to produce these long-term alterations is unknown.

Enduring cognitive effects of early malnutrition: a theoretical reappraisal.

This article presents a reappraisal of the literature on the enduring cognitive effects of early malnutrition. In addition to summarizing the existing empirical literature, we present a theoretical framework for determining whether the processes likely to be most vulnerable to early malnutrition were adequately assessed. The two types of information used to make this determination are clinical and experimental behavioral data as well as reported neural changes. One point of clear consensus is that animals exposed to early malnutrition exhibit lasting changes in the realm of emotionality, motivation, and/or anxiety. Because these alterations profoundly affect all aspects of behavioral functioning, including cognition, it is suggested that future research focus on these changes, rather than control for them as many past studies have done. The functional integrity of specific cognitive processes is less clear. The only cognitive processes for which enduring cognitive changes were demonstrated in rehabilitated animals--outside of effects mediated by these affective changes--are cognitive flexibility and, possibly, susceptibility to proactive interference. However, the inference that these are the only processes affected does not appear to be warranted on the basis of the evidence that several cognitive processes likely to be affected have not been fully assessed. Examples include executive functions linked to the prefrontal cortex (for example, attention), transfer of learning, procedural learning and long-term memory. Future research focusing on these specific cognitive functions as well as on these unequivocal affective changes should allow a more definitive conclusion regarding the enduring functional consequences of early malnutrition.

Accuracy and reliability of total body electrical conductivity (TOBEC) for determining body composition of rats in experimental studies.

Total body electrical conductivity (TOBEC) has been promoted as a noninvasive method to estimate body composition in small mammals. Validation of this method has primarily been under normative conditions and has generally been inadequate. This article reports on the reliability and accuracy of TOBEC methodology to assess gradual, physiologically induced changes in body composition in rats under different experimental conditions. Reliability of the index of electrical conductivity (EM number) was assessed by analyzing components of variance. Accuracy was assessed by comparing EM number to actual lean body mass (LBM, from carcass analysis), across different experimental conditions, within a particular experimental condition, and over time for a given set of animals. Reliable measurements were obtained by strictly adhering to a standard protocol. TOBEC was inaccurate across experimental conditions, within experimental conditions, and within a single experimental condition during the course of an experiment. This inaccuracy apparently stemmed from the lack of a direct relationship between EM number and LBM; EM number was more strongly correlated with body weight than with LBM. At the present time, TOBEC cannot be used in place of carcass analysis to accurately predict the body composition of rats during or following the administration of a variety of experimental conditions.

Chronic fenfluramine treatment: effects on body weight, food intake and energy expenditure.

Fenfluramine suppressess the body weight of experimental animals during chronic treatment by a mechanism that is not yet clear. The present research investigated the contributions of fenfluramine-induced alterations in food intake and two components of energy expenditure, resting energy expenditure (REE) and the thermic effect of food (TEF), to maintenance of a lowered body weight in male Long Evans rats. DI-Fenfluramine (20 mg/kg) produced an initial severe anorexia and weight loss. Food intake then increased steadily, reaching control levels by the fourth day of treatment. Tolerance to the anorectic effects of fenfluramine was not complete however -- food intake of fenfluramine-treated rats fell below control levels on 7 of the remaining 20 days of treatment. The body weight of fenfluramine treated rats remained significantly below control levels throughout the entire treatment period, but rose rapidly to control levels following fenfluramine withdrawal. REE was unaffected by fenfluramine treatment. By contrast, fenfluramine treatment significantly altered the TEF. Twenty-four hours after initiation of treatment the TEF of fenfluramine treated animals was higher than that of controls. On the 15th day of drug administration and the first day of fenfluramine withdrawal, however, the TEF of control and treated animals no longer differed. By contrast, on the second day of fenfluramine withdrawal the TEF of fenfluramine treated rats was suppressed to 17% of control levels. These findings suggest that the effect of fenfluramine on body weight involves modulation of both food intake and energy expenditure.

Deficient cumulative learning: an animal model of retarded cognitive development.

Biological insults that produce profound mental retardation (MR) in humans have generally been found to produce little cognitive dysfunction in animal models. Based on the fact that impaired transfer of learning is one of the hallmark characteristics of mentally retarded humans, we proposed that this discrepancy may largely reflect the common use of a single learning task as the critical cognitive index rather than an assessment of cumulative learning. Consistent with this hypothesis, rats exposed to prenatal hyperphenylalaninemia (a model of maternal PKU) evidenced significant impairment when tested on a series of 10 problems designed to allow for positive transfer of learning. This same treatment, however, did not alter learning rate of the individual tasks that comprised this series when presented singly to experimentally naive animals. Deficient transfer of learning contributed significantly to the impairment observed in the maternal PKU group. These results support the hypothesis that the assessment of cumulative learning is an important component of animal models of impaired cognitive development.